1. Field of the Invention
The present invention relates to a base body having micropores and a method of manufacturing the base body.
More specifically, the invention relates to a base body having fine vacuum holes that trap microorganisms or cell bodies and a method of manufacturing the base body.
In addition, more specifically, the invention relates to a base body having micropores that form a lipid membrane and a method of manufacturing the base body.
2. Description of the Related Art
As a method of physically trapping a cell or cultivated cell taken from a living organism and performing electrophysiological measurement on the cell, a patch-clamp method is known.
As a method of physically trapping a cell of the related art, a method of sucking a cell 101 at a hole provided on the side surface of a resin well 108 (FIG. 45) is known (see, Adrian Y. Lau, et al., Lab Chip, 2006, 6, 1510-1515 (Non Patent Document 1)).
Non Patent Document 1 discloses a method in which the behaviors of the cell trapped at the hole are directly observed using a high-power microscope from the rear surface of an apparatus and an apparatus that can perform electrophysiological measurements on the cell membrane and the like of the cell trapped at the hole.
The method and the apparatus become effective as means for explaining new biological characteristics.
Generally, in the patch-clamp method, there is a demand for closely attaching the adsorption portion (opening portion) at the hole and the cell membrane so as to achieve robust sealing of 1 gigaohm or more (high resistance sealing).
In order to form the above high resistance sealing, it is necessary to stabilize the sealing between the cell membrane and the adsorption portion, and a decrease in the diameter of the opening of the adsorption portion and a level difference or seam-free hole shape are required.
The cell trapping apparatus disclosed in Non Patent Document 1 has a base body to which two members formed of a resin (PDMS) are adhered, and micro holes are provided in the interface of the adhesion.
The base body is provided with a space in which a fluid is accumulated.
The space is formed by punching the base body.
When the base body is punched, the micro holes formed in advance in the base body are opened to the space.
Since the cell trapping apparatus is manufactured using a method of punching a resin, it is difficult to process the shape of the adsorption portion (opening portion) of the micro holes which is to trap a cell into a desired shape.
In addition, in a case in which a plurality of adsorption portions (opening portions) are formed in the base body, it is difficult to form the plurality of adsorption portions (opening portions) into a uniform shape.
In addition, since the base body is a resin, it is difficult to form fine holes having a minor axis of approximately less than 2 μm.
Furthermore, since the micro holes are formed by adhering two members to each other, a seam or level difference at which the two members are adhered to each other is present in the adsorption portion.
When the above seam or level difference is present, there is a case in which stable trapping of a cell is difficult.
That is, since the accuracy of the size or shape of the micro holes or the adsorption portion formed in the cell trapping apparatus is low, there is a problem in that it is difficult to trap a cell as desired.
In addition, in a case in which an electrophysiological measurement is conducted, since the sealing between the cell membrane and the adsorption portion becomes unstable, there is a problem in that a highly accurate electrophysiological measurement is difficult.
Furthermore, in order to trap microorganisms (having a short side on the order of nanometers) that are smaller than cells, it is necessary to highly accurately form the diameter of the short side of the adsorption portion (opening portion) of the micro holes at least on the order of nanometers.
However, since the cell trapping apparatus is configured by adhering the plurality of resin substrates 110 and 112 in Non Patent Document 1, it is difficult to process the adsorption portion (opening portion) on the order of nanometers.
Meanwhile, as a method of detecting a macromolecule such as a protein material included in a cell only at an extremely small amount, the western blotting of the related art or the method of Patent Document 1 (PCT International Publication No. WO2008-539711) is known.
However, while the movement distance of a protein or the like differs depending on the molecular weight, the movement distance may be as extremely short as approximately 100 nm to 350 nm.
Therefore, in the western blotting having a large volume of detection area, a large amount of cells are required in order for antibodies to come into contact with and trap an extremely small amount of a protein material.
Meanwhile, in the method of Patent Document 1, since the fluidic channel is provided in the side wall including PDMS, it is difficult to process the minor axis of the fluidic channel on the order of nanometers.
Therefore, there is a low probability that an extremely small amount of a protein material comes into contact with the analysis components (detection portion) of the fluidic channel having a minor axis on the order of micrometers, and a large amount of cells are required in order to obtain a desired amount of a protein material.
In addition, in the apparatus disclosed in Patent Document 1, since the fluidic channel is formed by adhering two members to each other, a seam or level difference at which the two members are adhered to each other is also present at the fluidic channel input end, and the sealing between a cell and the fluidic channel input end becomes unstable.
Therefore, when a cell is sucked, a void is caused between the cell and the fluidic channel, and there is a possibility that an extremely small amount of a protein material will not intrude into the fluidic channel and leaks outside.
As described above, in any method, a large amount of cells are required, and there is a problem in that detection is impossible unless a cell fragment is taken from a patient or the like.
In order to stably carry out an electrophysiological measurement and increase accuracy, decreasing the cross-section area of a hole that traps a cell may become effective.
Therefore, there is a demand for development of an apparatus having a hole with a diameter of the opening which is smaller than the diameter (approximately 2 μm to 4 μm) of the opening of a hole in the related art and a method of manufacturing the apparatus.
In addition, for the hole with a diameter of the opening which is smaller than the diameter (approximately 2 μm to 4 μm) of the opening of a hole in the related art, there is a demand for development of an apparatus that can reconfigure the same lipid bilayer membrane as the cell membrane artificially and a method of manufacturing the same.
In addition, as described above, there is a demand for development of an apparatus having micropores which can be used to trap fine particles such as microorganisms and cells, form a lipid membrane, and the like, and a method of manufacturing the apparatus.
On the other hand, in order to quantitatively detect a macromolecule such as a protein material which is included only a small amount of cell or the like efficiently, there is a demand for development of an apparatus which has nanofluidic channels, has no seam at the fluidic channel input end, and is formed of a transparent base member so that fluorescent labels can be observed, and a method of manufacturing the apparatus.